Effective Mass Spectrometer (MS)-Based Methods for Systematic and Unambiguous Characterization of Protein O-GlcNAcylation

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With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Ronghu Wu and his group at Georgia Tech are working to improve our ability to characterize an important group of natural protein modifications that involve attachment of sugars. Specifically, Dr. Wu is developing biotechnology tools based on mass spectrometry, in search of better understanding of O-GlcNAcylation, a protein modification which impacts cell function (for example, regulation of gene expression). Graduate and undergraduate students engaged in the research receive valuable training in instrumentation, chemical measurements, and large-scale data analysis. Dr. Wu also provides research opportunities for high school students and teachers, exposing them to cutting-edge research and stimulating their interests in STEM. The Wu group seeks to comprehensively and unambiguously characterize protein O-GlcNAcylation through the development of novel mass spectrometry (MS)-based methods. The problem is challenging because of the low natural abundance of the target proteins in complex biological samples, and the need to distinguish proteins with O-GlcNAc from those with stereoisomeric O-GalNAc. Dr. Wu is developing protocols which selectively enrich O-GlcNAcylated proteins, enabling global and quantitative analysis of protein O-GlcNAcylation. Identification of glycoproteins in the cytosol and the nucleus, and quantification of their dynamics, will provide valuable information to facilitate our understanding of protein O-GlcNAcylation, with potential applications in biological and biomedical research.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.